tbuckley/cabot-search huggingface dataset:
Abstract 5819: Src inhibitor suppresses rab11 mediated collective cell migration in colon cancer cells
Authors: Yi-Wen Lu, Xiang-Ling Hou, Hui Min Koo, Wei-Ting Chao
Journal: Cancer Research (2023)

Abstract Collective cell migration has been demonstrated the importance in cancer metastasis. Instead of single cell migration, collective cell migration requires E-cadherin for the cohort cell migration. Rab11 the recycling vesicle protein plays the key role in E-cadherin dynamics and mediates collective cell migration. In this study, Src was hypothesized to regulate Rab11 mediated collective cell migration. Our results demonstrated Src inhibitor Dasatinib has less toxicity in HT-29 colon cancer cells, instead, the cell morphology was turned aggregated. The immunofluorescence image also showed E-cadherin polarity was lost after Dasatinib treatment and increased the potential of cell-cell contact inhibition. In transwell assay, Rab11 was found colocalized with E-cadherin in collective migrated cells. Dasatinib treatment can suppress collective cell migration and decrease the interaction of Rab11 and E-cadherin. This study demonstrated the important role of Src in Rab11 mediated collective cell migration through E-cadherin dynamics in colon cancer. Citation Format: Yi-Wen Lu, Xiang-Ling Hou, Hui Min Koo, Wei-Ting Chao. Src inhibitor suppresses rab11 mediated collective cell migration in colon cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5819.
Abstract 4726: KMT5C-dependent regulation of mesenchymal-related genes and identification of KMT5C interactome using BioID
Authors: Jihye Son, Alejandra Agredo, Ally Glaws, Andrea L. Kasinski
Journal: Cancer Research (2023)

Abstract Epigenetic dysregulation has recently been recognized as a new hallmark of cancer. Dysregulation of chromatin remodelers is estimated to affect 10-20% of cancer. Thus, understanding how epigenetic-based mechanisms contribute to cancer initiation and progression is critical. KMT5C is a histone methyltransferase that catalyzes histone H4 lysine 20 trimethylation (H4K20me3), which has historically been associated with formation and maintenance of constitutive heterochromatin regions. Albeit recent evidence from our lab also suggests regulation of gene-rich euchromatin regions. In this previous body of work we discovered that loss of KMT5C promotes development of resistance to tyrosine kinase inhibitors (TKI) in EGFR-mutant non-small cell lung cancer (NSCLC) cells (PC9 and HCC827). Patient data further supports that the KMT5C transcript level is downregulated following resistance to osimertinib, a third-generation TKI. Our data suggests that KMT5C is involved in non-canonical regulation of genes outside of heterochromatic regions through its methyltransferase activity. Analysis of data deposited into the Cell Miner database and publicly available RNA-sequencing and ChIP-sequencing data revealed that the KMT5C-H4K20me3 axis has significant negative correlations with several genes involved in epithelial-to-mesenchymal transition (EMT). Our validation studies support this observation. However the mechanisms involved in KMT5C facilitating this change at EMT-related genes is unknown. KMT5C is well understood to be recruited to heterochromatin regions through its interaction with HP1, yet the mechanism for recruitment of KMT5C to gene-rich euchromatin regions remains to be elucidated. We hypothesize that KMT5C is recruited to its target genes in euchromatin in an HP1-independent mechanism. In this study, we aim to identify the interactome of KMT5C using an unbiased proximity labeling BioID approach. This study is expected to uncover a novel epigenetic mechanism that contributes to development of resistance to TKIs and may inform of strategies to target KMT5C-downregulated TKI-resistant cancers. Citation Format: Jihye Son, Alejandra Agredo, Ally Glaws, Andrea L. Kasinski. KMT5C-dependent regulation of mesenchymal-related genes and identification of KMT5C interactome using BioID. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4726.
Abstract 3868: Aurora kinase A inhibition overcome adaptive resistance to KRAS G12C inhibitor by G1-checkpoint induced apoptosis in KRAS non-small cell lung cancer
Authors: Chendi Li, Jeremy Chang, Mohammed U. Syed, Anahita Nimbalkar, Yi Shen, Steve Altschuler, Lani Wu, Xueqian Gong, Aaron Hata
Journal: Cancer Research (2023)

Abstract Cancer cells gain drug-tolerant states and evade therapy. In response to KRAS G12C inhibitor (G12Ci), KRAS mutant non-small cell lung cancer (NSCLC) cells maintain a drug-tolerant state by aurora kinase A (AURKA). AURKA can phosphorylate c-Raf to maintain new KRAS signaling, however, how AURKA becomes activated to cause KRAS G12Ci resistance is unclear. We show here that KRAS G12C + AURKA inhibition cause synthetic lethality in KRAS G12C NSCLC cells. LY3499446 (KRAS G12Ci) and LSN3321213 (aurora kinase A inhibitor) induced apoptosis that is independent of inhibited MAPK reactivation. Using high-content imaging that tracks single-cell fate during treatment, we observed that single-agent KRAS G12Ci induces G1 arrest in a sub-population of cells. Upon co-treatment with G12C + AURKAi, cells that are halted in G1 phase undergo early apoptosis, while those that initially evade G1 arrest and proceed through G2/M undergo apoptosis in subsequent G1. These data suggest the hypothesis that AURKA inhibition may increase the probability of G1 checkpoint-induced apoptosis by facilitating chromosomal misalignment and genomic instability. In summary, we provide clinical rationale for clinical testing of KRAS G12C + AURKA inhibitors. We also suggest a novel mechanism explaining the dependency of KRAS G12C resistant subpopulations on AURKA, leading to the opportunity to investigate the role of genomic instability in conferring KRAS G12Ci adaptive resistance. Citation Format: Chendi Li, Jeremy Chang, Mohammed U. Syed, Anahita Nimbalkar, Yi Shen, Steve Altschuler, Lani Wu, Xueqian Gong, Aaron Hata. Aurora kinase A inhibition overcome adaptive resistance to KRAS G12C inhibitor by G1-checkpoint induced apoptosis in KRAS non-small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3868.
Abstract 6237: Small molecule screen identifies pyrimethamine as an inhibitor of NRF2-driven esophageal hyperplasia
Authors: Chorlada Paiboonrungruang, Zhaohui Xiong, Yahui Li, Brittany Bowman, Zhi Huang, Vivian Chen, Bernard Weissman, Kevin P. Williams, M Ben Major, Xiaoxin Chen
Journal: Cancer Research (2023)

Abstract Objective: NRF2 is a master transcription factor that regulates the stress response. NRF2 is frequently mutated and activated in human esophageal squamous cell carcinoma (ESCC), which drives resistance to chemotherapy and radiation therapy. Therefore, a great need exists for NRF2 inhibitors for targeted therapy of NRF2high ESCC. Design: We performed high-throughput screening of two compound libraries from which hit compounds were further validated in human ESCC cells and a genetically modified mouse model. Mechanism of action of one compound was explored by biochemical assays. Results: Using high-throughput screening of two small molecule compound libraries, we identified 11 hit compounds as potential NRF2 inhibitors with minimal cytotoxicity at specified concentrations. We then validated two of these compounds, pyrimethamine and mitoxantrone, by demonstrating their dose- and time-dependent inhibitory effects on the expression of NRF2 and its target genes in two NRF2Mut human ESCC cells (KYSE70 and KYSE180). RNAseq and qPCR confirmed the suppression of global NRF2 signaling by these two compounds. Mechanistically, pyrimethamine reduced NRF2 half-life by promoting NRF2 ubiquitination and degradation in KYSE70 and KYSE180 cells. Expression of an Nrf2E79Q allele in mouse esophageal epithelium (Sox2CreER;LSL-Nrf2E79Q/+) resulted in an NRF2high phenotype, which included squamous hyperplasia, hyperkeratinization, and hyperactive glycolysis. Treatment with pyrimethamine (30mg/kg/day, p.o.) suppressed the NRF2high esophageal phenotype with no observed toxicity. Conclusion: We have identified and validated pyrimethamine as an NRF2 inhibitor that may be rapidly tested in the clinic as a radiation and chemotherapy sensitizer for NRF2high ESCC. Citation Format: Chorlada Paiboonrungruang, Zhaohui Xiong, Yahui Li, Brittany Bowman, Zhi Huang, Vivian Chen, Bernard Weissman, Kevin P. Williams, M Ben Major, Xiaoxin Chen. Small molecule screen identifies pyrimethamine as an inhibitor of NRF2-driven esophageal hyperplasia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6237.
Abstract 6772: Powerful background reduction in fluorescent tissue stains with an improved proximity-based technology for detection of protein-protein interactions
Authors: Hampus Elofsson, Doroteya Raykova, Agata Zieba Wicher
Journal: Cancer Research (2023)

Abstract Immunofluorescent staining of tissues via in situ proximity ligation assay (isPLA) is now a well-established tool for highly sensitive detection of protein-protein interactions and their localization. It finds versatile applications in basic research, understanding of cellular processes, visualization of tissue architecture, and identification of biomarkers, among others. However, standard IF and isPLA techniques alike may suffer from high background. Tissues are particularly affected by background problems caused by autofluorescence and tissue infiltration of various cell types (e.g., immune, endothelial) where the fluorescently labelled oligos used for detection in isPLA may bind unspecifically. To address this, we developed a next generation isPLA-based technology for fluorescent detection of protein interactions in FFPE and fresh frozen human and mouse tissues, called NaveniFlex Tissue. In contrast to previously available protocols, it can visualize signal that would otherwise be obscured by background, thereby increasing sensitivity. Here, we compare the ability of NaveniFlex Tissue and similar commercial kits to detect the interaction between low abundance proteins Podocalyxin and Ezrin in the glomeruli of healthy kidney and in breast cancer, where co-expression is a prognostic marker for increased metastatic potential. While other kits generate high background which masks isPLA signal entirely, NaveniFlex Tissue dramatically reduces cell-based background and visualizes the interaction in its specific localization. Furthermore, in a TMA staining for the Mucin 16/Mesothelin interaction, our method successfully demonstrates strong and clear signal in stage III ovarian cancer. Moreover, it sensitively detects even low abundance interaction in a stage Ia tumor. In contrast, a different commercial kit detects significantly less interactions in the stage III tumor, while in the lower grade tumor it does not generate signals above background level. Therefore, NaveniFlex Tissue improves detection in tissues with varying disease progression, thus adding information and prognostic value to the staining. Taken together, our data illustrate that NaveniFlex Tissue outperforms current isPLA-based techniques by efficiently reducing background, which improves visualization of signal and signal-to-noise ratios in various healthy and diseased tissues. Citation Format: Hampus Elofsson, Doroteya Raykova, Agata Zieba Wicher. Powerful background reduction in fluorescent tissue stains with an improved proximity-based technology for detection of protein-protein interactions. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6772.
Abstract 3867: Chromatin modification driving sub-clonal resistance to KRAS G12C combination therapies in KRAS mutant non-small cell lung cancer
Authors: Chendi Li, Qian Qin, Mohammed Usman Syed, Anahita Nimbalkar, Barbara Karakyriakou, Sarah E. Clark, Anne Y. Saiki, Paul E. Hughes, Chris Ott, Luca Pinello, Aaron N. Hata
Journal: Cancer Research (2023)

Abstract The FDA approval of the KRAS G12C inhibitor (G12Ci) sotorasib and the advancement of similar drugs into clinical trials marks a major milestone in treating KRAS G12C non-small cell lung cancer (NSCLC). However, not all patients respond (sotorasib - ORR = 37.1%, adagrasib - 43%, JDQ443 - 35%), motivating preclinical and clinical investigation into mechanisms of intrinsic and acquired resistance. For instance, clinical studies have reported on-target KRAS mutations and preclinical studies have demonstrated mitogen-activated protein kinase (MAPK) feedback reactivation including EGFP, SHP2, and WT RAS signaling. In response to targeted therapies, sub-populations of cells can enter quiescence or specific epigenetic-driven states that confer drug tolerance. However, epigenetic states defining drug-tolerant persister populations and contributing to adaptive resistance to KRAS G12Ci have not been reported. Using a lineage tracing barcoded system, we identify distinct and reversible subpopulations defined by specific chromatin and transcriptional states in KRAS NSCLC cell lines that contribute to KRAS G12Ci resistance in vitro, even prior to drug treatment. We observed that specific states, including activation of histone demethylation and SWI/SNF complex, may contribute to MAPK reactivation-driven resistance. These results suggest potential epigenetic vulnerabilities that can be exploited to improve the response to KRAS G12Ci. Moreover, we observed distinct persister subpopulations with resistance to KRAS G12Ci combination co-targeting orthogonal pathways (SHP2, CDK4/6, PI3K, and MCL-1), raising the possibility that distinct epigenetic-transcriptional states contribute to differential drug response and clonal evolution of persisters. Collectively, these results suggest that more complete tumor regression may be achieved by orthogonal strategies that target different resistant populations within the same tumor. Citation Format: Chendi Li, Qian Qin, Mohammed Usman Syed, Anahita Nimbalkar, Barbara Karakyriakou, Sarah E. Clark, Anne Y. Saiki, Paul E. Hughes, Chris Ott, Luca Pinello, Aaron N. Hata. Chromatin modification driving sub-clonal resistance to KRAS G12C combination therapies in KRAS mutant non-small cell lung cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3867.
Abstract 671: Exploiting macrophage differentiation and plasticity as an immunomodulating strategy
Authors: Carla Castro, Philipp Metzger, Cynthia Obodozie, Holger Weber
Journal: Cancer Research (2023)

Abstract The infiltration of macrophages into the tumor microenvironment (TME) is a well-known characteristic of tumor progression and therapy resistance. Depending on the signals derived from TME, macrophage plasticity allows them to undergo a variety of polarization states. Two extremes of macrophage polarization are M1 and M2 phenotypes. M1 macrophages provide enhanced anti-tumor inflammatory reaction while M2 macrophages are usually associated with tumor progression due to their immune suppression, angiogenesis and neovascularization promoting capabilities. Modulating the macrophage differentiation or polarization has emerged as a therapeutic strategy to synergize or complement current treatments. At Reaction Biology we have established human macrophage assay pipelines to evaluate the novel macrophage polarization compounds in vitro. Both, monocyte cell lines like THP-1, and and freshly isolated monocytes from donor derived PBMCs, are of great value. Donor derived differences could become an issue during the screening phase compared to the use of cell lines. Yet, after the screening phase, the evaluation across several healthy donors is crucial to assess the impact of a compound on the general population. Monocytes from primary human peripheral blood mononuclear cell (PBMC) are isolated, differentiated into naïve macrophages, and polarized. Effects of compounds modulating the differentiation and polarization of macrophages are analyzed by flow cytometry. Moreover, the functional reprogramming of monocytes and macrophages can be analyzed by measuring the phagocytic or tumor cell killing capacity via flow cytometry or using high content screening equipment. In the addition, a wide range of secreted cytokine can be quantified by MSD ELISA. Our data shows that human primary monocytes grown under M1 polarizing conditions express high levels of CD86 and HLA-DR in contrast to those cultivated under M2 polarizing conditions, with high expression of CD163 and CD206. A pH-sensitive phagocytosis could be shown for M2 macrophages. Modulation of surface markers from polarized macrophages under different treatment conditions could be observed, as well as changes in their functionality, supporting the translational value to evaluate the efficacy of immunomodulating compounds in vitro. Citation Format: Carla Castro, Philipp Metzger, Cynthia Obodozie, Holger Weber. Exploiting macrophage differentiation and plasticity as an immunomodulating strategy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 671.
Abstract 4215: Air pollution exposure during pregnancy and risk of premenopausal breast cancer among women with a family history of cancer
Authors: Anna Hettinger, Jessica Edlund, Malin Gustafsson, Jenny Lindén, Wendy Yi-Ying Wu, Anna Oudin, Sophia Harlid
Journal: Cancer Research (2023)

Abstract Background: Having a family history of breast or ovarian cancer is an important risk factor for premenopausal breast cancer. One reason for this could be that women with an underlying genetic susceptibility are more sensitive to environmental exposures due to gene-environment interactions. This might be especially important if the exposure occur during windows of susceptibility, such as pregnancy. Aim: The aim of this project is to determine if exposure to air pollution during pregnancy is associated with premenopausal breast cancer in a population with a known family history of breast or ovarian cancer. Methods: The project is based on nation-wide data from Sweden’s national registers and utilizes a cohort study design. The original cohort consists of women who gave birth to their first child between 1992 and 2015 (n=1 074 925), from this cohort we will select women with at least one first degree relative with breast or ovarian cancer (confirmed through register linkage). The primary outcome is primary breast cancer diagnosed before 50 years of age, and the exposure is levels of air pollution (PM10, PM2.5 and NO2) at the participant’s home address during each pregnancy. Cox proportional hazards regression will be used to determine hazard ratios using age as the time-scale. Results: The registry based linkage makes it possible to distinguish between both relational context (mother or sister), number of afflicted first degree relatives and type of cancer (breast or ovarian). All included women also have at least one recorded pregnancy (prior to any cancer diagnosis). Conclusion: This large scale study will be able to provide important information about the impact of air pollution exposure during pregnancy on breast cancer risk in a population with a documented family history of breast or ovarian cancer. Citation Format: Anna Hettinger, Jessica Edlund, Malin Gustafsson, Jenny Lindén, Wendy Yi-Ying Wu, Anna Oudin, Sophia Harlid. Air pollution exposure during pregnancy and risk of premenopausal breast cancer among women with a family history of cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4215.
Abstract 1890: Bispecific T cell engagers targeting TrkB
Authors: Jetta Bijlsma, Srinidhi Desikan, Timothy Miller, Allie Thom, Jennifer Bath, Ilse Roodink
Journal: Cancer Research (2023)

Abstract Tropomyosin kinase B (TrkB) plays an important role in survival, differentiation, and proliferation of neural cells. It belongs to the family of receptor tyrosine kinases and its specific ligands are NT-4/5 and BDNF. The (over)expression of TrkB and (over)activation of the BDNF/TrkB signaling pathway is implicated in various solid malignancies and associated with poor prognosis due to the role of TrkB in suppressing anoikis and promoting epithelial-mesenchymal transition. Here we present the development and characterization of novel T cell engagers targeting TrkB. A panel of 41 sequence unique antibodies were selected from human phage display libraries and B cells of TrkB-ECD immunized chicken. Following recombinant expression, the panel of antibodies was analyzed for off-rate, binding reactivity to plate-immobilized and cell-expressed TrkB, cross-reactivity to other tropomyosin kinase family members, and competition with BDNF. Based on their characteristics, two targeting arms were prioritized for the generation of T cell engagers using a standard bispecific knob-into-hole approach. Apart from a commercial CD3 targeting arm, three proprietary CD3 targeting arms were also analyzed. After successful production of the bispecifics , the molecules were validated to bind to plate-immobilized TrkB and CD3 separately and cell-associated TrkB and CD3 simultaneously. In addition, our TrkB T cell engagers were able to induce varying levels of T cell activation in vitro. Our data suggests that these TrkB/CD3 bispecific molecules could be interesting candidates for further development into immuno-oncology therapeutics. Citation Format: Jetta Bijlsma, Srinidhi Desikan, Timothy Miller, Allie Thom, Jennifer Bath, Ilse Roodink. Bispecific T cell engagers targeting TrkB [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1890.